Identification and characterization of a third thioredoxin h in poplar

Three full-length sequences encoding thioredoxin h have been isolated in a leaf/root of Populus trichocarpa cv. Trichobel expressed sequence tag (EST) library. One of these, popCXXS1 exhibits the nontypical active site CXXS homologous to atCXXS1. The second one, named popTrxh4, is related to atTrxh9...

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Bibliographic Details
Published in:Plant physiology and biochemistry 2003-06, Vol.41 (6), p.629-635
Main Authors: Gelhaye, Eric, Rouhier, Nicolas, Vlamis-Gardikas, Alexios, Girardet, Jean-Michel, Sautière, Pierre-Eric, Sayzet, Michel, Martin, Francis, Jacquot, Jean-Pierre
Format: Article
Language:English
Subjects:
Biochemistry, Molecular Biology
CHEMICOPHYSICAL PROPERTIES
COMPOSÉ ORGANOSOUFRÉ
COMPUESTO ORGÁNICO DEL AZUFRE
Life Sciences
Medicin och hälsovetenskap
MUTANT
MUTANTES
MUTANTS
NUCLEOTIDE SEQUENCE
ORGANOSULPHUR COMPOUNDS
OXIDOREDUCTIONS
OXIRREDUCIÓN
OXYDORÉDUCTION
POPULUS
POTENCIAL REDOX
POTENTIEL REDOX
PROPIEDADES FISICOQUÍMICAS
PROPRIÉTÉ PHYSICOCHIMIQUE
PROTEINS
PROTEÍNAS
PROTÉINE
RECOMBINACIÓN
RECOMBINAISON
Recombinant
RECOMBINATION
REDOX POTENTIAL
Redox regulation
SECUENCIA NUCLEOTÍDICA
SÉQUENCE NUCLÉOTIDIQUE
Thioredoxin h
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Summary:Three full-length sequences encoding thioredoxin h have been isolated in a leaf/root of Populus trichocarpa cv. Trichobel expressed sequence tag (EST) library. One of these, popCXXS1 exhibits the nontypical active site CXXS homologous to atCXXS1. The second one, named popTrxh4, is related to atTrxh9 which forms with several other plant thioredoxin h a distinct subgroup of thioredoxins h. The third one, named popTrxh3, displays 66% identity and also a high degree of homology (81%) vs. the previously described popTrxh1. Nevertheless, the active sites of both proteins differ, since the active site of popTrxh1 (WCPPC) is a variant of the canonical WCGPC found in popTrxh3. The cDNA sequence of popTrxh3 has been introduced in an expression plasmid (pET3d) in order to express the corresponding recombinant polypeptide. The protein has been expressed to a high level, purified from Escherichia coli cells with a high yield and its catalytic properties compared to popTrxh1. Furthermore, two mutants, popTrxh1P40G and popTrxh3G41P, have been engineered in order to explore the importance of the active site residues in the thioredoxin h catalytic properties. The results are discussed in relation with known biochemical properties of thioredoxins h.
ISSN:0981-9428
1873-2690
DOI:10.1016/S0981-9428(03)00063-9